Anchoring of heterologous proteins in multiple Lactobacillus species using anchors derived from Lactobacillus plantarum

Abstract

Members of the genus Lactobacillus have a long history in food applications and are considered as promising and safe hosts for delivery of medically interesting proteins. We have assessed multiple surface anchors derived from Lactobacillus plantarum for protein surface display in multiple Lactobacillus species, using a Mycobacterium tuberculosis hybrid antigen as test protein. the anchors tested were a lipoprotein anchor and two cell wall anchors, one non-covalent (LysM domain) and one covalent (sortase-based anchoring using the LPXTG motif). Thus, three different expression vectors for surface-anchoring were tested in eight Lactobacillus species. When using the LpXtG and LysM cell wall anchors, surface display, as assessed by flow cytometry and fluorescence microscopy, was observed in all species except Lactobacillus acidophilus. Use of the cell membrane anchor revealed more variation in the apparent degree of surface-exposure among the various lactobacilli. Overproduction of the secreted and anchored antigen impaired bacterial growth rate to extents that varied among the lactobacilli and were dependent on the type of anchor. overall, these results show that surface anchors derived from L. plantarum are promising candidates for efficient anchoring of medically interesting proteins in other food grade Lactobacillus species.

Surface-Displayed Amuc_1100 From Akkermansia muciniphila on Lactococcus lactis ZHY1 Improves Hepatic Steatosis and Intestinal Health in High-Fat-Fed Zebrafish

“…Lactococcus lactis ZHY1 competent cells were prepared and transformed as described previously with slight modifications ( 11 ). The overnight culture of L. lactis was inoculated at a 5% ratio into a fresh GM17 medium with 2.5% glycine, and then cultivated until OD 600 = 0.5.…”

Lactiplantibacillus plantarum as a Potential Adjuvant and Delivery System for the Development of SARS-CoV-2 Oral Vaccines

“…Although most of them are L. lactis -based vaccines, the immunogenicity of L. plantarum -based vaccines was reported to be significantly higher than that of L. lactis [ 129 ]. Since recombinant L. plantarum expressing heterologous antigens can induce both specific humoral and cellular immune responses against pathogens and toxin in mucosal tissues [ 129 , 130 , 131 , 132 , 133 , 134 , 135 , 136 ] and the oral administration of L. plantarum can effectively stimulate antiviral immune responses [ 87 , 88 ], during past decades, L. plantarum have been evaluated as vehicles for delivering recombinant viral antigens [ 131 , 137 , 138 ].…”

“…To date, several delivery systems based on L. plantarum were established for cell-surface expression of viral antigens ( Table 1 ) [ 137 , 138 , 139 ]. A cell wall anchoring system for E7 mutant protein (E7mm) of human papillomavirus (HPV) type-16 was constructed by fusing the signal peptide and the first 15 amino acids of lactococcal Usp45 protein at the N terminal of the viral protein, and the cell wall anchor of L. plantarum lp_2940 protein at the C terminal [ 139 ].…”

“…The E7mm protein can be efficiently anchored and displayed on the cell wall of L. plantarum via anchor motif LPQTXE [ 139 ]. However, the display efficiency of heterologous protein on the cell surface may be affected by signal peptide, surface anchor and host cell surface structure [ 137 , 139 , 140 ]. As for L. plantarum , the heterologous antigen can be displayed on the surface of the bacteria via C-terminal or N-terminal anchors.…”

“…The cell-surface display efficiency of three surface anchors in eight different Lactobacillus were evaluated and it was found that LPXTG- and LysM-based anchors of L. plantarum exhibited higher efficiency than that of the lipoprotein anchor [ 137 ]. Notably, heterogenicity was observed in antigen display cells and cell populations.…”